Abstract: Objective　To understand the kdr (knockdown resistance, kdr) gene mutation of the voltage-gated sodium channel (VGSC) of Anopheles sinensis in Yunnan Province. Methods　From 2018 to 2019, mosquitoes were collected in Luoping County, Suijiang County, Tengchong City, Yingjiang County, Yuanjiang County and Mengla County in Yunnan Province. The collected mosquitoes were morphologically identified as Anopheles sinensis and genomic DNA was extracted by kits. The DNA templates were sequenced after PCR amplification and the sequencing results were identify as Anopheles sinensis by homology alignment in NCBI. After the ⅡS5 and ⅡS6 fragments of the sodium channels in Anopheles sinensis were amplified and sequenced, the sequencing results were multiple aligned by DNAMAN software, and the mutations were analyzed one by one with BioEdit software to determine the kdr allele types and genotypes, and the frequencies were calculated. Results This survey amplified 287 sequences, and the sequence maps showed that 1014 loci had three alleles, including wild type TTG/L (89.20%), mutant type TTT/F (9.76%) and TCG/S (1.04%). Five genotypes: homozygous wildtype L/L (85.02%), homozygous mutant F/F (6.27%) and S/S (0.35%), heterozygous mutant L/F (6.97%) and L/S (1.39%). The wild type allele TTG/L was the main allele in six sampling sites except Suijiang County. The frequency of wild type allele in Tengchong City was the highest (100.00%). That is, no mutation was detected, while the rest of counties occurred different degrees of mutation at 1014 loci. The frequency of mutant allele in Suijiang County was the highest, reaching 55.68%. Luoping County, Mengla County and Suijiang County had two mutant types. Yingjiang County and Yuanjiang County had one heterozygous mutant L/F. Conclusion　Wild type L1014 (TTG/L) is still dominant in most areas of Yunnan Province. The kdr mutation type is mainly L1014F, followed by L1014S, and the mutation frequency is lower than that in central provinces of China.

Objective: To construct a virtual reconstruction method including midspan maxillary defects and provide clinical reference by training a generative adversarial network (GAN) model. Methods: The CT data of middle-aged Han patients with oral diseases who visited the Department of Radiology, West China Hospital of Stomatology, Sichuan University from June 2015 to June 2022 were collected, where the CT data of 100 healthy maxilla and 15 maxillary defects (5 simple unilateral defects, 5 unilateral defects involving zygomatic bone, 5 midspan defects) were selected. Mimics was used to create spherical phantom and simulate bone defects around the healthy maxillas, including simple unilateral defects, unilateral defects involving zygomatic bone and midspan defects. The original image was set as the correct reference for the reconstruction: artificial defects paired with the correct reference were divided into training set (n=70), validation set (n=20) and test set (n=10), where the first two were used to train the GAN model, and the test set was used to evaluate the GAN performance. Data from 15 clinical defects were imported into the trained GAN model for reconstruction, with mirroring and GAN-based virtual reconstruction for unilateral clinical defects, and only the latter method was adopted for midspan defects. The reconstruction results were divided into mirror reconstruction group (n=10), unilateral defect GAN reconstruction group (n=10) and midspan defect GAN reconstruction group (n=5). The test set, mirror reconstruction group, and unilateral defect GAN reconstruction group were quantitatively evaluated, whose quantitative indicators were Dice similarity coefficient (DSC) and 95% Hausdorff distance (HD95), and the group results were subjected to one-way ANOVA and Tukey test. The test set, mirror reconstruction group, unilateral defect GAN reconstruction group and midspan defect GAN reconstruction group were qualitatively scored, and Kruskal-Wallis test and Bonferroni correction were used for the total score of each group. Results: The total differences in the test set, mirror reconstruction group, unilateral defect GAN reconstruction group DCS (0.891±0.049, 0.721±0.047, 0.778±0.057, respectively) and HD95 [(3.58±1.51), (5.19±1.38), (4.51±1.10) mm, respectively] were statistically significant (F=28.08, P<0.001; F=3.62, P=0.041); among them, the test set DSC was significantly larger than the mirror reconstruction group (P<0.05), and the test set HD95 was significantly less than the mirror reconstruction group (P<0.05). Overall difference in qualitative total scores [8 (1), 6 (2), 6 (2), and 4 (2) points, respectively] in the test set, mirror reconstruction group, unilateral defect GAN reconstruction group, and midspan defect GAN reconstruction group were statistical significance (H=18.13, P<0.001); pairwise comparison showed that the total score of the test set was significantly higher than that of the mirror reconstruction group (P<0.05). Conclusions: The virtual reconstruction method based on GAN proposed in this study has better virtual reconstruction effect of unilateral defect than mirror technique, and can also realize virtual reconstruction of maxillary midspan defect.
Humans ; Middle Aged ; Maxilla/surgery* ; Deep Learning ; China

Professor
Acupuncture ; Acupuncture Therapy ; Angina, Stable ; Combined Modality Therapy ; Humans ; Moxibustion

To learn the current situation and strengthen the management of national standards for Chinese medicinal materials, we sorted out the relevant national standards. According to incomplete statistics, there are 1 185 kinds of Chinese medicinal materials, including 1 024 kinds of plant medicines, 106 kinds of animal medicines, and 54 kinds of mineral medicines, in addition to ethnic medicinal materials with different functions. The relevant standards include 819 Pharmacopoeia standards, 342 standards issued by the Ministry of Health or National Medicinal Products Administration, 7 standards for new medicinal materials, and 17 standards for imported medicinal materials. In this paper, the sources of standards as well as the distribution of families and genera and the distribution of medicinal parts of medicinal materials are analyzed. The suggestions are as follows:(1)to improve the coordination among different national standards of Chinese medicinal materials;(2)to improve the standardization and controllability of relevant standards;(3)to revise the issued standards for Chinese medicinal materials(including Tibetan, Uygur, and Mongolian medicinal materials).
Animals ; Asians ; China ; Drugs, Chinese Herbal ; Humans ; Medicine, Chinese Traditional ; Plants, Medicinal ; Reference Standards

OBJECTIVE: To observe the effect of electroacupuncture (EA) combined with lifestyle control on hepatic fat status, hepatic enzymology, glycolipid metabolism and anthropological parameters in patients with obese nonalcoholic fatty liver disease(NAFLD). METHODS: A total of 90 patients with obese NAFLD were randomized into an observation group (45 cases, 4 cases dropped off) and a control group (45 cases, 1 case dropped off). Lifestyle control was implemented in the control group. On the basis of the treatment in the control group, acupuncture was applied at Zhongwan (CV 12), Quchi (LI 11), Shuifen (CV 9), Huaroumen (ST 24), Daheng (SP 15), Guanyuan (CV 4), Qihai (CV 6), etc. EA was provided at Huaroumen (ST 24) and Daheng (SP 15) with dilatational wave, 2 Hz/100 Hz in frequency, 30 min each time, once every other day, 3 times a week. The treatment for 12 weeks was required in both of the two groups. Hepatic fat status [controlled attenuation parameter (CAP) and liver stiffness measurement (LSM)], hepatic enzymology [alanine aminotransferase (ALT), aspartate aminotransferase (AST) and γ-glutamyl transferase (GGT)], glycolipid metabolism and insulin sensitivity [fasting plasma glucose (FPG), fasting serum lisulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C)] and anthropological parameters [body weight (BW), body mass index (BMI), fat percentage (FP), waist circumference (WC), hip circumference (HC) and waist-to-hip ratio (WHR)] in the two groups were observed before and after treatment. RESULTS: ①Compared before treatment, hepatic CAP, LSM, serum ALT, AST and GGT after treatment were obviously reduced in the two groups (<0.05, <0.01). After treatment, CAP and ALT in the observation group were lower than the control group (<0.05). ②Compared before treatment, FINS, HOMA-IR, LDL-C, TC and TG after treatment were obviously reduced in the two groups (<0.05, <0.01),while the levels of HDL-C were increased (<0.05). Compared before treatment, FPG after treatment in the observation group was reduced (<0.05). Compared with the control group, FINS, HOMA-IR, TC and TG in the observation group were lower than those in the control group after treatment (<0.05). ③Compared before treatment, BW BMI, FP, WC, HC, WHR after treatment were obviously reduced in the two groups (<0.01). After treatment, WC and WHR in the observation group were lower than the control group (<0.05). CONCLUSION Electroacupuncture combined with lifestyle control can effectively treat obese nonalcoholic fatty liver disease, and present better therapeutic effect on hepatic fat status, glycolipid metabolism, insulin resistance, WC and WHR.

Objective Sclerosing pneumocytoma (SP) is a rare benign tumor of the lung. This study investigates the CT and PET/CT features of 52 patients with sclerosing pneumocytoma (SP), summarizes their imaging features and provide reference for improving the accuracy of SP imaging diagnosis.Methods The clinical and imaging data of 52 patients with SP confirmed by operation and pathology in the Affiliated Hospital of Nanjing University of Traditional Chinese Medicine and the First Affiliated Hospital of Nanjing Medical University from January 2010 to March 2018 were analyzed retrospectively，and the imaging features of the disease were analyzed and summarized. Among 52 SP patients, 46 were female and 6 were male, with an average of 50.7±12.6 years old.Results ①Morphological characteristics: all were round or round-shaped solitary nodules or masses in the lung. The average diameter of the tumor was 22.2±15.4 mm. 39 lesions had smooth edges and 13 had shallow edge lobulation sign; ②Predilection sites: 33 of the cases were closely attached to or adjacent to the pleura, and there was no preponderant distribution of the lobe; ③Density: The density of the lesion was homogeneous. 8 cases had coarse calcifications at the edges or in the lesions; ④Enhancement features: 26 cases showed moderate to marked enhancement, only 6 cases were mild enhancement; ⑤Features around the tumor: "vascular adhering sign" in 21 cases, "tail sign" in 7 cases, "air crescent sign " in 5 cases, "localized emphysema" in 4 cases, "halo sign" in 11 cases, "bird egg-bird nest sign" in 5 cases, 4 cases of secondary bronchiectasis; ⑥Associated tumors: 3 cases of invasive lung adenocarcinoma, 3 cases of lung adenocarcinoma in situ, 2 cases of pulmonary atypical adenomatous hyperplasia, 1 case of pulmonary tiny inflammatory nodule; 2 cases of esophageal cancer associated, 1 case of invasive thymoma, 1 case of large mature ovarian cystic teratoma; ⑦PET/CT characteristics: 18F-FDG standard uptake value in tumor increased to different degrees, the average of SUV max value was 4.4±3.2.Conclusion The innovation of this research: SP is shown as solitary circular or circular-shaped blood-rich nodules in the lung, often under the pleura, especially the mediastinal pleura and lobar pleura. There is no lobar distribution advantage; SP calcification is uncommon, among which the calcification at the margin of the tumor was more common. "Air crescent sign", " Halo sign" and " Bird egg-bird nest sign" have high diagnostic value for SP.

Objective To investigate the epidemiological and molecular virological characteristics in HBV-infected patients with copositive HBsAg and anti-HBs.Methods HBV serological markers were analyzed in 52 070 specimens.The epidemiological characteristics of HBsAg and anti-HBs simultaneously positive patients (the experimental group) and HBsAg positive and auti-HBs negative patients (the control group) were compared.The S protein of HBV coding region was amplified by semi-nested PCR and sequenced.The statistical differences between the two groups were compared in different gene regions,genotypes and different clinical diagnosis.Results HBsAg was positive in 20.40％ (10 621/52 070) of all specimens.In the patients with positive HBsAg,2.48％ (263/10 621) was positive anti-HBs.The prevalence of co-positive HBsAg and auti-HBs was higher in aged 0 to 9 years and greater than or equal to 80 years than that in other age,and the prevalence of positive HBsAg and negative anti-HBs was completely opposite.The mutation rate of S protein in the experimental group was significantly higher than that in the control group (1.52％ vs 0.81％,P ＜0.01) with the mutation in the major hydrophilic region (MHR) (1.68％ vs 0.57％,P ＜0.01).The mutation rates of S protein of HBV carriers,chronic hepatitis B (CHB) patients and patients with liver cirrhosis (LC) in the experimental group were significantly higher than those in the control group (1.47％ vs 0.65％,1.28％ vs 0.84％,2.21％ vs 0.44％,P ＜0.05,respectively),except for the patients with hepatocellular carcinoma (HCC) (1.97％ vs 2.21％,P ＞ 0.05).Conclusion Co-positive HBsAg and anti-HBs in HBV-infected patients was more common in HBsAg positive patients aged 0 to 9 years and greater than or equal to 80 years than the others.Coexistence of HBsAg and anti-HBs in HBV-infected patients may relate to immune escape caused by mutation of S protein (mainly MHR).The mutation rates of S protein in the two groups of patients,co-positive HBsAg and anti-HBs and the positive HBsAg combined with negative anti-HBs,were associated with the stage of liver disease.

[Objective]To establish a reliable and accurate preimplantation genetic diagnosis (PGD)method using multiple dis?placement amplification (MDA), which can be applied to the diagnosis of X-linked severe combined immunodeficiency disease (X-SCID).[Methods]Haplotype analysis for the X-SCID family was performedusing five short tandem repeats (STR) markers flanking the both sides of the interleukin-2 (IL-2) receptor gamma chain (IL2RG) gene. MDA technique was used for single-cell whole genomic amplification. The products were used as template in polymerase chain reaction (PCR) of informative STR markers found by linkage analysis for haplotype analysis as well as sequencing of the IL2RG gene exon 5.The amelogenin (AMEL) locus was used to do sex diag?nosis.[Results]Linked analysis revealed 3 STR markers were informative. The method was evaluated with 10 single lymphocytes and 10 single blastomeres. MDA was successful in all single cell. The detection efficiency of gene sequencing of pathogenic IL 2RG exon5 was 100%. The PCR efficiency of 3 STR informative markers and AMEL was 96.3%(77/80)and the average allele drop-out (ADO) rate was 11.5%(7/61). A cycle of PGD was performed on the family, and seven embryos were diagnosed, two of which were normal embry?os. Twin pregnancy occurred after transplantation which were given a healthy baby boy and a healthy baby girl.[Conclusion]In this study, multiple displacement amplification combined with specific amplification/sequencing of pathogenic gene and haplotype analysis in the single cell level of X-linked severe combined immunodeficiency disease were performed. The protocol can avoid misdiagnosis caused by contamination and ADO, and improve the diagnostic efficiency of PGD.

OBJECTIVETo evaluate the clinical outcomes between extreme lateral interbody fusion and conventional posterior operation in the treatment of upper lumbar disc herniation.

METHODSAmong 60 patients with upper lumbar disc herniation were treated with extreme lateral interbody fusion(XLIF) or conventional posterior operation from June 2010 to December 2014, 30 patients(19 males and 11 females) were treated with XLIF (XLIF group); and the other 30 patients(17 males and 13 females) were treated with conventional posterior operation (conventional group). In XLIF group, the lesions occurred at T₁₂L₁ segments in 2 patients, at L₁,₂ segments in 6 patients, at L₂,₃ segments in 10 patients, and at L₃,₄ segment in 12 patients. In conventional group, the lesions occurred at T₁₂L₁ segments in 1 patient, at L₁,₂ segments in 6 patients, at L₂,₃ segments in 8 patients, and at L₃,₄ segment in 15 patients. Operative incision lengths, time, blood loss, postoperative draining volume, hospital stays were recorded. Pre-and post-operative visual analogue score(VAS) and Japanese Orthopedic Association(JOA) were compared between two groups. According to the image data, the intervertebral fusion device was observed to be displaced and the rate of interbody fusion was analyzed.

RESULTSAll the patients were followed up, and the duration ranged from 12 to 48 months, with an average of 29 months. The complications included 2 femoral nerve damage in XLIF group (postoperative recovery within 3 months) and superficial incision infection in conventional group(cured by anti-infection). There were no patients with cerebrospinal fluid leakage(CSFL), cauda equina injuries or functional deterioration in the nerve root of lower limbs. In the XLIF group: the operative time was (65.6±20.5) minutes, blood loss was (48.8±15.3) ml, postoperative draining volume was 0 ml. In the conventional group: the operative time was (135.2±33.9) minutes, blood loss was (260.3±125.7) ml, postoperative draining volume was (207.1±50.2) ml. The operative time, blood loss, postoperative draining volume in XLIF group were less than those in the conventional group(<0.05). The JOA and VAS score were significantly improved in both groups during the follow-up period compared with those before operation(<0.05). But the difference of the JOA and VAS score between the two groups 1, 6, and 24 months after surgery had not significant differences(>0.05). There were no significant differences in the fusion rate between the two groups 6 and 12 months after operation(>0.05).

CONCLUSIONSThe XLIF fusion for the treatment of upper lumbar disc herniation has several advantages such as minimal invasive, stable vertebral plate, less complications and postoperative fusion rate, which has a better clinical effect.

OBJECTIVEIn this study, milk from a cow with mastitis was analyzed to determine the presence of mycobacterial infection. Milk quality and security problems pertaining to the safe consumption of dairy products were also discussed in this study.

METHODSMilk was preprocessed with 4% NaOH. Then, mycobacteria were isolated from the milk sample on L-J medium. The isolate was identified using multiple loci Polymerase Chain Reaction (PCR) and multi-locus sequence analysis with 16S rRNA, sodA, hsp65, and ITS genes. The drug sensitivity of the isolate to 27 antibiotics was tested through alamar blue assay.

RESULTSSmooth, moist, pale yellow colonies appeared on the L-J medium within a week after inoculation. Based on the results of multiple loci PCR analysis, the isolate was preliminarily identified as non-tuberculous mycobacteria. The 16S rRNA, SodA, hsp65, and ITS gene sequences of the isolate exhibited 99%, 99%, 99%, and 100% similarities, respectively, with those of the published reference strains of Mycobacterium elephantis (M. elephantis). The drug sensitivity results showed that the strain is resistant to isoniazid, p-aminosalicylic acid, and trimesulf but is sensitive to ofloxacin, rifampicin, amikacin, capreomycin, moxifloxacin, kanamycin, levofloxacin, cycloserine, ethambutol, streptomycin, tobramycin, rifabutin, ciprofloxacin, linezolid, cefoxitin, clarithromycin, and minocycline.

CONCLUSIONTo the best of our knowledge, this study is initially to report the isolation of M. elephantis from the milk of a cow with mastitis in China.

Animals ; Anti-Bacterial Agents ; pharmacology ; Cattle ; China ; Drug Resistance, Bacterial ; Female ; Mastitis, Bovine ; epidemiology ; microbiology ; Milk ; microbiology ; Mycobacterium ; drug effects ; genetics ; isolation & purification ; Mycobacterium Infections ; epidemiology ; microbiology ; veterinary ; Phylogeny ; Polymerase Chain Reaction